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Method for producing cadaverine dicarboxylate

a technology of cadaverine and dicarboxylate, which is applied in the field of cadaverine dicarboxylate production, can solve the problems of inability to meet high temperature use, inconvenient production methods of biomass, complex process, etc., and achieve the effect of simple and efficient production

Active Publication Date: 2005-01-06
AJINOMOTO CO INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides a method for economically producing cadaverine as a raw material for production of nylon in a form easily and most efficiently used in the polymerization. This is achieved by subjecting a lysine solution to an enzymatic decarboxylation reaction and maintaining the pH of the solution at a level sufficient for the reaction to occur by adding dicarboxylic acid to the solution. The method can also produce a cadaverine dicarboxylate which can be used in a polymerization reaction for producing nylon.

Problems solved by technology

However, since polylactic acid has a melting point of about 190° C., it is not suitable for high temperature uses.
However, production methods from biomass are unknown.
Therefore, the process becomes complicated and is no longer economical.

Method used

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  • Method for producing cadaverine dicarboxylate

Examples

Experimental program
Comparison scheme
Effect test

example 1

Construction of a LDC-Amplified Strain of Escherichia coli

[0065] PCR primers having the nucleotide sequences of SEQ ID NOS: 1 and 2 were designed based on the nucleotide sequence of the LDC gene (cadA) of E. coli (Watson N. et al., Journal of Bacteriology, Vol. 174, 530-540, 1992; Meng S. Y. and Bennet G. N., Journal of Bacteriology, Vol. 174, 2659-2669, 1992) and used for PCR with the chromosome of E. coli W3110 (ATCC 39936) as a template to amplify a DNA fragment containing the cada gene.

[0066] To prepare the pcadA plasmid, the amplified DNA fragment was digested with KpnI and SphI, and the resulting fragment (2468 bp) was inserted into the KpnI-SphI digestion site of pUC18 (Takara Shuzo) (FIG. 1). The E. coli JM109 strain (Takara Shuzo) was transformed with the pcadA plasmid. A transformant was selected using ampicillin resistance as a marker, and designated E. coli JM109 / pcadA.

example 2

Production of Cadaverine Adipate from Lysine Adipate Using cadA-Amplified Strain

[0067] (1) Culture of cadA-Amplified Strain

[0068]E. coli JM109 / pcadA was precultured in the LB medium, and then 50 ml of the culture broth was inoculated into 500 ml of the LB medium of two-fold concentration (2% of trypton, 1% of yeast extract, 1% of NaCl) contained in a 1-L jar fermenter (ABLE Co., Ltd.). The cells were cultured with aeration and stirring under the following conditions: aeration rate of 250 ml / min, 35° C. and 700 rpm. Following a 15 hour culture, the whole culture broth was inoculated into 22 L of LB medium of two-fold concentration contained in a 50-L jar fermenter, and the culture was continued under the following conditions: aeration rate of 11 L / min, 35° C., an internal pressure in the jar of 50 kPa and 250 rpm. Following the 4 hour culture, 3 g of IPTG (isopropyl-β-D-thiogalactopyranoside) was dissolved in 50 ml of water and added to the culture through a filter. Then, the cultu...

example 3

Acquisition of Cadaverine Adipate Crystals

[0075] (1) Removal of cells from cadaverine adipate solution

[0076] The cadaverine adipate solution obtained in Example 2 was sterilized in an autoclave at 120° C. for 10 minutes and centrifuged to collect a supernatant.

[0077] (2) Decoloration and Concentration

[0078] The obtained supernatant was added to 20% activated carbon based on the cadaverine weight and decolored with stirring at 20° C. for 1 hour. The activated carbon was removed using filter paper, and the obtained filtrate was concentrated 4- to 5-fold under reduced pressure (55 to 60° C., 110 to 150 mmHg). The solid content of the concentrate was 70 to 77%.

[0079] (3) Crystallization of Cadaverine Adipate and Separation of Crystals

[0080] The aforementioned concentrate was cooled from 60° C. to 10° C. at 4° C. / hour to precipitate crystals. The crystallization rate was 40 to 45%. The precipitated crystals were separated and collected using a centrifuge and air-dried in a desiccat...

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Abstract

Cadaverine dicarboxylate is produced by performing an enzymatic decarboxylation reaction of a lysine solution while adding a dicarboxylic acid containing 4 to 10 carbons to the lysine solution to maintain pH of the solution at a level sufficient for the enzymatic decarboxylation reaction to occur, for example, 4.0 to 8.0.

Description

BACKGROUND OF THE INVENTION [0001] 1. Field of the Invention [0002] The present invention relates to a method for producing cadaverine dicarboxylate. Cadaverine dicarboxylate can be used as a raw material for producing nylon. [0003] 2. Description of the Related Art [0004] Naphtha, a fossil material, is a major raw material in the production of plastics. Disposal of plastics which are not recycled has recently become a environmental concern due to the release of carbon dioxide when disposed of by burning, etc.. Accordingly, with the goal of preventing global warming and encouraging a recycling society, it is strongly desirable to replace raw materials for producing plastics with those derived from biomass. [0005] Polylactic acid is known as a plastic produced using biomass as a raw material. A method for producing polylactic acid includes first extracting starch or sugar from a plant, then producing lactic acid by fermentation using the extracted starch or sugar as a carbon source, ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C08G69/26C12N1/21C12P13/00
CPCC08G69/26C12P13/02C12P13/001
Inventor NISHI, KIYOHIKOENDO, SHUICHIMORI, YUKIKOTOTSUKA, KAZUHIKOHIRAO, YOSHINORI
Owner AJINOMOTO CO INC
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